• 제목/요약/키워드: T-세포

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시험관 내 면역 기술에 의한 항체 생산에 관한 연구

  • 전태훈;서동상
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.19-32
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    • 1994
  • 가장 효과적인 H-2b 항원에 대한 시험관내 항체 생산 조건을 찾기 위하여 근교계 생쥐인 C57BL/6BySnj의 비장세포를 UV로 불활성학 시킨 후 항원으로 사용하고, A/wySnJ$\times$Sm/J(ASmJF1, hybrid)의 비장세포를 항원 수용자 계통으로 하여 5-7일동안 배양기에서 항체 생산을 유도하였다. 본 실험에서 T 임파구 대식세포와 임파구 분화 촉진 인자인Concanavalin A Lipopolvsaccharide. Pokeweed mitogen 등을 사용하여 20가지 조건으로 실험을 수행하여, 항체 생성 여부는 보체 의존성 세포 장애 실험과 면역 효소법에 의해 조사하였다 그 결과 모든 조건에서 항체생산이 확인되었으며. 가장 좋은 시험관내 항체 생산 조건으로는 T 임파구와 대식세포를 함께 사용하여 면역시킨 것이 가장 효과적이었다. 이 방법을 이용하여 항체 생산을 유도한 후 5일째 면역된 비장세포를 Sp2/0-Ag 14와 세포 융합시켜 H-2b 마우스의 체포 표면 항원에 대한 단일군항체 생산을 시도하였다. 또한 생체내 면역 방법과 비교하기 위해 6주간 C57BL/6BySnJ의 비장세포를 복강내에 주사하여 같은 조건으로 세포융합을 시도하였다. 그 결과 H-2b 세포의 표면 항원에 대한 항체 생산을 하는 세포군은 시험관내 면역 방법에서 3개 생체내 면역 방법에서 4개부 확인되었다.

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Anti-Obesity Effects of Red Onions Extract (적양파 추출물의 항비만 활성)

  • Song, Hwan;Seo, Ji-Hun
    • Journal of Convergence for Information Technology
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    • v.12 no.3
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    • pp.126-131
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    • 2022
  • Obesity is known as a metabolic disease caused by abnormal differentiation of fat tissue due to an imbalance between energy intake and consumption.. The purpose of this study was to confirm the changes in the genes associated with pancreatic lipase activity and pre-adipocyte cell differentiation by treatment of red onion extract treatment. The effect of red onion extract treatment on pre-adipocyte differentiation was evaluated using 3T3-L1 adipocytes, and the activity of related genes was confirmed through Real-Time PCR. As a result of the experiment, the red onion extract inhibit pancreatic lipidase activity by concentration dependent manner. In addition, it was found to inhibit adipocyte differentiation and inhibit the activity of genes(C/EBP-α, C/EBP-β, PPAR-γ) associated with adipocyte differentiation. Through the results of this experiment, it is suggested that the red onion extract can be developed as a high potential material with anti-obesity efficacy by suppressing adipocytic differentiation by controlling genes related to adipocyte differentiation.

Effects of PGA-LM on CD4+CD25+foxp3+ Treg Cell Activation in Isolated CD4+ T Cells in NC/Nga Mice (NC/Nga 생쥐에서 분리한 T 세포에서 foxp3+ 세포 활성화에 대한 PGA-LM의 효과)

  • Jang, Soon-Nam;Kim, Kum-Lan;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.160-169
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    • 2009
  • Poly-$\gamma$-glutamic acid ($\gamma$-PGA) was mixed natural flora of Bacillus subtilis, contaminated from cooked soybeans. Also, it was performed to find out the antiallergic activity by using NC/Nga mice, in vitro. The $\gamma$-PGA (PGA-HM : PGA-high molecular weight), Molecular weight 300 kDa, was decomposed and made PGA-LM (PGA-low molecular weight) which has molecular weight below 30 kDa by sonication. Therefore, it was same result between PGA-HM and PGA-LM, and reported PGA-LM as basic result. We found that PGA-LM contains antiallergic efficacy that inhibit B cells and Th2 cells activation from isolated CD4+T cells in NC/Nga atopic dermatitis model mice, and not show a cytotoxicity in the hFCs. To investigate the effects of these PGA-LM in vitro, isolation of splenic B cell and CD4+ T cells in atopic dermatitis mice were used. To elucidate the role of PGA-LM in anti-CD40+ interleukin-4 (IL-4)-mediated B-cell activation, showed that the capacity of B cells to expression IL-$1\beta$, IL-6, and TNF-$\alpha$ mRNA down-regulated, and IL-10 mRNA up-regulation by PGA-LM treatment, but it had no effect on TGF-$\beta$ expression. In addition to CD4+IFN-$\gamma$+ and CD4+CD25+foxp3+, the functions of PGA-LM in the development of the CD4+CD25+foxp3+ and CD4+IFN-$\gamma$+cells, the phenotype and functions of PGA-LM induced CD4+CD25+foxp3+, and CD4+IFN-$\gamma$+cells in CD4+T cells. These results suggested that PGA-LM could change cytokine production and generate CD4+CD25+foxp3+ Tregs in NC/Nga mice, and may be effective for immunotherapy in patients with AD.

Enhancement of Immune Activities of Ganoderma lucidum Mycelium Cultured with Garlic Enriched Medium (마늘 첨가 복합배지에서 배양된 영지 균사체의 면역 증진 효과)

  • Mun, Hyoung-Chul;Lee, Hyun-Soo;Park, Jin-Hong;Kim, Dae-Ho;Lee, Shin-Young;Seong, Nak-Sul;Bang, Jin-Ki;Jung, Hae-Gon;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.1
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    • pp.24-30
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    • 2004
  • The immune activities of Ganoderma lucidum Mycelium added garlic extracts (GAM), Ganoderma lucidum Mycelium (GM), garlic extracts (GS) and standard $({\beta}-glucan)$ were compared. GAM enhanced the growth of human immune T cell up to $1.25{\sim}1.46$ times, compared to control group. GAM showed relatively lower cytotoxicity in using normal human lung cell, while GAM showed the most potent inhibitory effect on the human lung carcinoma, compared to GM and GS. The selectivity of GAM was also higher than that of GM and GS. GAM increased the secretion of cytokines, IL-6 and TNF- from human B cell as well as the growth of human immune cells. It can imply that GAM has higher immune activity than GM or GS.

Steap4 Stimulates Adipocyte Differentiation through Activation of Mitotic Clonal Expansion and Regulation of Early Adipogenic Factors (Steap4에 의한 지방세포분화 촉진 기전)

  • Sim, Hyun A;Shin, Jooyeon;Kim, Ji-Hyun;Jung, Myeong Ho
    • Journal of Life Science
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    • v.30 no.12
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    • pp.1092-1100
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    • 2020
  • The six-transmembrane epithelial antigen of prostate 4 (Steap4) is a metalloreductase that plays a role in intracellular iron and cupper homeostasis, inflammatory response, and glucose and lipid metabolism. Previously, Steap4 has been reported to stimulate adipocyte differentiation; however, the underlying mechanisms of this action remain unexplored. In the present study, we investigated the molecular mechanisms involved in Steap4-induced adipocyte differentiation using 3T3-L1 cells, immortalized brown adipocyte (iBA) cells, and mouse embryonic fibroblast C3H10T1/2 cells. The knockdown of Steap4 using adenovirus-containing shRNA attenuated mitotic clonal expansion (MCE), as evidenced by the impaired proliferation of 3T3-L1 cells, iBA cells, and C3H10T1/2 cells within 48 hr after adding the differentiation medium. Steap4 knockdown downregulated G1/S phase transition-related cell cycle regulators (including cyclin A and cyclin D) and upregulated cell cycle inhibitors (including p21 and p27). Furthermore, Steap4 knockdown inhibited the phosphorylation of p38 mitogen-activated protein kinase, extracellular signal-regulated kinase, and Akt. Moreover, Steap4 knockdown repressed the expression of early adipogenic activators, such as CCAAT-enhancer-binding protein β (C/EBPβ) and Kruppel-like factor family factor 4 (KLF4). On the other hand, Steap4 knockdown stimulated the expression of adipogenic inhibitors, including KLF2, KLF3, and GATA2. The overexpression of Steap4 using an adenovirus removed the repressive histone marks H3K9me2 and H3K9me3 on the promoter of C/EBPβ. These results indicate that Stepa4 stimulates adipocyte differentiation through the induction of MCE and the modulation of early adipogenic transcription factors, including C/EBPβ, during the early phase of adipocyte differentiation.

Ethanol Extract of Hippophae Rhamnoides L. Leaves Inhibits Adipogenesis through AMP-activated protein kinase (AMPK) Activation in 3T3-L1 Preadipocytes (비타민나무 잎 에탄올추출물의 AMPK 활성화를 통한 3T3-L1 지방전구세포의 adipogenesis 억제효과)

  • Jeong, Hyeon Ju;Park, Ju Hee;Kim, Myong-Jo
    • Korean Journal of Plant Resources
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    • v.28 no.5
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    • pp.582-590
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    • 2015
  • In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

Effects of Raphani Semen Ethanol Extracts on Skin Inflammation in HaCaT Keratinocytes (나복자(萊菔子) 에탄올 추출물이 HaCaT 피부각질형성세포에서 피부염증 감소에 미치는 영향)

  • Kim, Keun-Lip;Hong, Chul-Hee;Lee, Kyou-Young
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.35 no.2
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    • pp.13-27
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    • 2022
  • 목적 : 본 연구는 인간피부각질형성세포(HaCaT keratinocytes) 모델을 TNF-α와 IFN-γ로 자극하여 내복자(萊菔子)의 피부염증 감소 및 만성 염증성 질환에 미치는 영향을 알아보고자 하였다. 방법 : 내복자(萊菔子) 에탄올 추출물(RSE)이 세포생존율에 미치는 영향을 확인하기 위하여 MTT assay를 시행하였다. 또한 RSE가 TNF-α와 IFN-γ로 자극한 HaCaT 세포에서 p-IκBα, p-ERK, p-JAK2, p-STAT1, p-STAT6의 발현과 periostin, TSLP 단백질 발현에 미치는 영향을 확인하였다. 결과 : RSE는 200㎍/㎖ 이하에서 세포 독성을 보이지 않았고, HaCaT keratinocytes에서 TNF-α와 IFN-γ자극에 의하여 증가된 IκBα, ERK의 인산화를 억제하였다. 또한 JAK2와 STAT1, STAT6의 인산화를 억제하였으며, periostin과 TSLP의 발현을 감소시켰다. 결론 : RSE는 HaCaT keratinocytes에서 pro-inflammatory cytokines 및 transcription factors의 발현을 감소시켜 피부염증 감소 효능을 보였고, 만성 염증성 질환에서 내복자(萊菔子)의 사용 가능성을 확인하였다.

Effects of Ethanol Extracts from Commonly Consumed Vegetables in Korea on Differentiation and Secretion of MCP-1 and Adiponectin in 3T3-L1 Adipocytes and Lipid Accumulation in HepG2 Hepatocytes (한국인 다소비 채소의 에탄올 추출물이 3T3-L1 지방세포와 HepG2 간장세포의 지질축적, MCP-1과 Adiponectin의 분비에 미치는 영향)

  • Ahn, Eun Mi;Kang, Hyun Ju;Kim, Young;Choe, Jeong Sook;Kang, Min-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.25 no.1
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    • pp.99-110
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    • 2015
  • The purpose of this study was to evaluate the inhibitory effects of commonly consumed vegetables in Korea on lipid accumulation and production of pro-inflammatory cytokines related to obesity/metabolic syndrome. Using KNHANES (Korea National Health and Nutrition Examination Survey) raw data ($1^{st}$; 1998, $5^{th}$; 2010, 2011) and a literature search, we selected vegetables for study. Edible portions of samples were prepared, ethanol-extracted, and then freeze-dried. 3T3-L1 adipocytes and HepG2 hepatocytes cells were used as in vitro models. Lipid accumulation determined by Oil-red O staining showed that all samples except bracken had inhibitory effects on lipid accumulation in 3T3-L1 adipocytes. Especially, crown daisy and mugwort effectively reduced accumulation of lipids, and their inhibition rates were more than 60% of the control group. Young pumpkin, leeks, crown daisy, and mugwort showed significantly decreased MCP-1 levels compared to the control group. However, adiponectin protein level did not increase in the vegetables experimental group. In HepG2 hepatocytes, all samples showed inhibitory effects on lipid accumulation at one of the two concentrations. Although adiponectin protein levels did not increase, MCP-1 protein levels decreased in adipocytes. Further, lipid accumulation in adipocytes and hepatocytes decreased. In conclusion, all samples showed one or more improved obesity/metabolic syndrome indicators. Among them, young pumpkin, leeks, crown daisy, and mugwort were selected as the most effective portions of vegetables based on improvement of obesity/metabolic syndrome-related indicators.

The Effect of A Static Magnetic Field on The Bone Nodule Formation of MC3T3-E1 Cells (정자기장이 MC3T3-E1 세포의 골결절형성에 미치는 영향)

  • Kim, Kwang-Duk;Kwon, Oh-Won;Ryu, Hyun-Mo
    • The korean journal of orthodontics
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    • v.28 no.1 s.66
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    • pp.155-163
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    • 1998
  • To evaluate the effect of a static magnetic field on the bone producing potential of MC3T3-E1 cells, the alkaline phosphatase activity was measured after the cells having been cultured under 76.4mT static magnetic field using a $SmCo_5$ magnets for 5days, 7days, 11days, 15days and 21days for each cell culture group. Also, the amount of bone nodule stained with Alizarin red S was observed. The results were as follows . The alkaline phosphatase activity of the 7, 11, and 15 days group among the experimental groups was decreased as compared with the control groups, and the decrease of alkaline phosphatase activity in the 11 days group was the most evident among them. . Any stained bone nodules of both groups had not been observed until the 11th day. The stained bone nodules in the control groups were found on the 15th day, but not in the experimental groups. The stained bone nodules were observed in both groups on the 21st day, but the control groups have more bone nodules than the experimental groups.

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Anti-Obesity Effect of Ethyl Acetate Extracts from Agrimonia pilosa Ledeb. in 3T3-L1 Preadipocytes (3T3-L1 지방전구세포에서 용아초 에틸아세테이트 추출물의 항비만 효과)

  • Lee, Jung-A;Ahn, Eun-Kyung;Hong, Seong-Su;Oh, Joa-Sub
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.2
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    • pp.161-167
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    • 2012
  • To evaluate the anti-obesity effect of Agrimonia pilosa L., this study investigated that ethyl acetate extract from A. pilosa L. (EAAP) suppresses lipid accumulation and inhibits expression of adipogenic marker genes, such as peroxisome proliferator activated receptor ${\gamma}$ (PPAR${\gamma}$), CCAAT-enhancer-binding protein ${\alpha}$ (C/EBP${\alpha}$), glucose transporter 4 (GLUT4), and adiponectin in 3T3-L1 preadipocytes. We demonstrated that EAAP inhibited adipocyte differentiation and expression of PPAR${\gamma}$ and C/EBP${\alpha}$ mRNA levels in a dose-dependent manner. In addition, EAAP reduced the PPAR${\gamma}$ transcriptional activity stimulated by rosiglitazone in HEK 293T cells and decreased the expression of GLUT4 and adiponectin in 3T3-L1 cells. These results suggest that EAAP inhibits preadipocyte differentiation and adipogenesis by blocking of PPAR${\gamma}$ and C/EBP${\alpha}$ gene expression in 3T3-L1 cells.